Suzuki H, Kamata T, Onishi H, Watanabe S
J Biochem. 1982 May;91(5):1699-705. doi: 10.1093/oxfordjournals.jbchem.a133861.
In 1978, we (Suzuki et al., J. Biochem. 84, 1529) reported the very interesting finding that in a medium of 0.2 M KCl (and 10 mM MgCl2), addition of ATP induced a large increase in the sedimentation velocity of chicken gizzard myosin from approximately 6S to 10S. Moreover, our flow birefringence study suggested that 10S-myosin was not much different from 6S-myosin in the particle length. Therefore, we concluded that ATP induced dimerization of gizzard myosin monomers. In the present study, we reinvestigated 6S-myosin and 10S-myosin by the sedimentation equilibrium method, and found that both myosins had the same molecular weight of approximately 500,000. We also studied the angular dependence of the light scattering intensity, and found that addition of ATP caused a large change in the radius of gyration of gizzard myosin; the radius of gyration of 6S-myosin was 545 A whereas that of 10S-myosin was only 146 A. Accordingly, our previous conclusion had to be withdrawn. Instead, we now put forward a new conclusion that ATP induces a large change in the conformation of gizzard myosin monomers. We added two new observations: (a) The ATP-induced change in the myosin conformation and the large decrease in the ATPase activity of myosin were both reversible upon increasing the KCl concentration from 0.2 M to 0.3 M. (b) The large decrease in the ATPase activity and the ATP-induced increase in the sedimentation velocity were also observed with gizzard heavy meromyosin when the KCl concentration decreased to lower than 0.3 M.
1978年,我们(铃木等人,《生物化学杂志》84卷,1529页)报道了一个非常有趣的发现:在含有0.2M氯化钾(和10mM氯化镁)的介质中,添加ATP会使鸡砂囊肌球蛋白的沉降速度大幅增加,从约6S增加到10S。此外,我们的流动双折射研究表明,10S肌球蛋白在颗粒长度上与6S肌球蛋白没有太大差异。因此,我们得出结论,ATP诱导了砂囊肌球蛋白单体的二聚化。在本研究中,我们通过沉降平衡法对6S肌球蛋白和10S肌球蛋白进行了重新研究,发现两种肌球蛋白的分子量相同,约为500,000。我们还研究了光散射强度的角度依赖性,发现添加ATP会导致砂囊肌球蛋白的回转半径发生很大变化;6S肌球蛋白的回转半径为545 Å,而10S肌球蛋白的回转半径仅为146 Å。因此,我们不得不撤回之前的结论。相反,我们现在提出一个新的结论,即ATP会使砂囊肌球蛋白单体的构象发生很大变化。我们补充了两个新的观察结果:(a)当氯化钾浓度从0.2M增加到0.3M时,ATP诱导的肌球蛋白构象变化和肌球蛋白ATP酶活性的大幅降低都是可逆的。(b)当氯化钾浓度降至低于0.3M时,砂囊重酶解肌球蛋白也观察到ATP酶活性的大幅降低和ATP诱导的沉降速度增加。